MACHEREY-NAGEL RNA purification guide Bioanalysis RNA purification products Subhead for various Cover sample materials n Tailored solutions for various applications n Superior reproducibility and quality n Highest lysis efficiency
MACHEREY-NAGEL RNA Kit Finder Target molecule Starting material Technology Format Product Page Micro NucleoSpin RNA XS p. 4 Mini NucleoSpin RNA p. 3 NucleoSpin RNA Plus p. 3 Midi NucleoSpin RNA Midi p. 3 Cells and tissue 8 / 96-well NucleoSpin 8/96 RNA p. 5 Reagent Scalable NucleoZOL p. 4 RNA (> 2 nt) Magnetic beads Scalable NucleoMag 96 RNA p. 5 Blood Mini NucleoSpin RNA Blood p. 9 Midi NucleoSpin RNA Blood Midi p. 9 8 / 96-well NucleoSpin 8 / 96 RNA Blood p. 9 Plant Mini NucleoSpin RNA Plant p. 11 Reaction mixtures Mini NucleoSpin RNA Clean-up p. 5 Micro NucleoSpin RNA Clean-up XS p. 5 Cells and tissue Mini NucleoSpin mirna p. 6 Reagent Scalable NucleoZOL p. 4 Small and large RNA (incl. mirna) NucleoZOL lysates Mini NucleoSpin RNA Set for NucleoZOL p. 4 Plasma Mini NucleoSpin mirna Plasma p. 6 FFPE Mini NucleoSpin totalrna FFPE p. 1 Micro NucleoSpin totalrna FFPE XS p. 1 RNA, DNA, and proteins Cells, tissue, plant, blood, bacteria, yeast Buffer set Mini Micro / Mini NucleoSpin TriPrep p. 7 NucleoSpin RNA/Protein p. 8 NucleoSpin RNA/DNA Buffer Set p. 8 Poly(A) mrna Total RNA Affinity beads Mini NucleoTrap mrna Mini p. 11 Midi NucleoTrap mrna Midi p. 11 2
NucleoSpin RNA Plus The next generation kit for RNA isolation from cells and tissue n NucleoSpin gdna Removal Column No time-consuming rdnase digestion n Lysate clearing and gdna removal with one column in a single step Covenient handling and very fast processing n New efficient lysis buffer No β-mercaptoethanol, TCEP, or other reducing reagents necessary 5 Time (min) 4 3 NucleoSpin RNA Plus Time saving RNA isolation 2 1 LT S P Z Q NS RNA The prep time of the NucleoSpin RNA Plus procedure was compared to the protocols of competitor kits (LT, S, P, Z, Q) as well as the prep time of the NucleoSpin RNA procedure (NS RNA) (Light grey: time for rdnase digestion). The NucleoSpin RNA Plus kit is the fastest RNA isolation kit on the market: high quality RNA in 2 minutes! NucleoSpin RNA Plus 1 / 5 / 25 74984.1 /.5 /.25 NucleoSpin RNA Mini spin kit for RNA isolation of highest integrity n Up to 7 μg ready-to-use RNA from different sample materials Cells, tissue, bacteria, yeast n rdnase included for on-column DNA digestion Easy and efficient removal of contaminating DNA n NucleoSpin Filters (shredders) Efficient sample homogenization and reduction of viscosity n For larger amounts of sample material (up to 2 mg tissue): NucleoSpin RNA Midi Cycle number 35 3 28.7 28.8 25 27.5 2 23.9 15 1 x 1 5 HeLa cells 5 mg mouse spleen NucleoSpin RNA Competitor Q Highly efficient removal of genomic DNA Total RNA isolated from different samples with NucleoSpin RNA and Competitor Q kits was analyzed by qpcr. The RNA was tested for residual traces of DNA by amplifying a 14 bp multi-copy target (HeLa cells) and a 191 bp multi-copy target (mouse spleen). Higher cycle numbers indicate less residual DNA. NucleoSpin RNA 1 / 5 / 25 74955.1 /.5 /.25 NucleoSpin RNA Midi 2 74962.2 3
NucleoZOL The universal RNA isolation reagent n No chloroform, no phase separation High RNA yields by a quick and easy procedure n High RNA purity from any sample material and any sample amount Efficient removal of DNA, proteins, and other contaminating compounds n Combination with NucleoSpin technology possible For even more convenient handling 12 Relative RNA yields (%) 1 8 6 4 2 mouse brain adipose tissue NucleoZOL=1% buffy coat Competitor Zol yeast wheat leaves Market-leading RNA yields RNA was extracted from different starting materials. RNA was quantified by qrt-pcr and relative yields were calculated (NucleoZOL=1 %). RNA isolation with NucleoZOL results in similar or better RNA yield compared to standard two-phase extraction methods (competitor Zol). Product Preps / Preps REF NucleoZOL 2 ml 7444.2 NucleoSpin RNA Set for NucleoZOL* 1 / 5 7446.1 /.5 * NucleoZOL reagent not included NucleoSpin RNA XS Purification of highly concentrated RNA from very small samples n Achieve higher sensitivities for downstream applications Optimized column design elution in as little as 5 μl possible n Successful RNA isolation even from minute sample amounts Ideal for biopsies, LCM samples, FACS sorted cells, single cells, fine needle aspirates n High-quality RNA, ready-to-use for RT-PCR and other applications Optimized procedure for highest RNA quality convenient and efficient on-column DNA removal 1 4 1 3 1 2 1 Fluorescence negative control 5 1 15 2 25 3 Cycle Number Reliable sensitivity constant high yields Total RNA was extracted from 1 4 (black), 1 3 (blue), 1 2 (green) and 1 (red) HeLa cells using NucleoSpin RNA XS. Elution was performed with 1 μl, of which 2 μl were subjected to a one-tube, one-step qrt-pcr. NucleoSpin RNA XS allows efficient isolation from sample amounts over several orders of magnitude, down to few cells. NucleoSpin RNA XS 1 / 5 / 25 7492.1 /.5 /.25 4
NucleoSpin RNA Clean-up NucleoSpin RNA Clean-up XS Mini und Micro kit for RNA clean-up n RNA clean-up from pre-purified RNA Phenol / chloroform extractions, enzymatic reactions, diluted RNA samples n Time-saving procedure without DNase digestion and homogenization steps Complete removal of RT-PCR inhibitors n Choose the format you need: NucleoSpin RNA Clean-up: For samples up to 2 µl reaction mixtures NucleoSpin RNA Clean-up XS: For very diluted samples up to 3 µl containing max. 9 µg RNA NucleoSpin RNA Clean-up 1 / 5 / 25 74948.1 /.5 /.25 NucleoSpin RNA Clean-up XS 1 / 5 / 25 7493.1 /.5 /.25 NucleoSpin 8 / 96 RNA RNA isolation in flexible 8-well strip or proven 96-well plate format n Time-saving parallel isolation of RNA from a variety of starting materials E.g., cells, tissue, plant, buffy coat, PAXgene Blood RNA Tubes n High-quality RNA rdnase included for efficient removal of genomic DNA MN Wash Plate minimizes risk of cross-contamination n Processing under vacuum or by centrifugation Suitable for manual and automated processing n NucleoSpin 8 / 96 RNA Core Kits Kits with basic content focused on automation platforms additional accessories can be combined as needed NucleoSpin 8 RNA 12 x 8 / 6 x 8 74698 /.5 NucleoSpin 8 RNA Core Kit 48 x 8 74465.4 NucleoSpin 96 RNA 2 x 96 / 4 x 96 / 24 x 96 7479.2 /.4 /.24 NucleoSpin 96 RNA Core Kit 4 x 96 74466.4 NucleoMag RNA Magnetic-bead based isolation of RNA n Reliable purification of high-quality RNA rdnase and reducing agent TCEP included Small elution volumes > 5 µl n Easily adaptable for automated use One-tube procedure minimizes risk of cross-contamination Complete processing at room temperature NucleoMag RNA 1 x 96 / 4 x 96 74435.1 /.4 5
NucleoSpin mirna Mini spin kit for isolation of small RNA, large RNA, and proteins n RNA purification with optional size selection Isolation of small RNA only (< 2 nt), small RNA and large RNA (> 2 nt) in two separate fractions, or total RNA (small and large RNA in one fraction) n Additional isolation of proteins from the same sample Ready to use for SDS-PAGE and Western blot analysis n Excellent RNA recovery and purity by chaotropic salt lysis without phenol / chloroform rdnase and NucleoSpin Filters included A: Total RNA [FU] 7 6 5 4 3 2 1 Marker peak 2 25 3 35 4 45 5 55 6 65 [s] [FU] 6 5 4 3 2 1 B: Small RNA fraction [FU] 6 4 2 Marker peak 2 3 4 5 6 [s] 2 25 3 35 4 45 5 55 6 65 [s] C: Large RNA fraction [FU] 6 5 4 3 2 1 Marker peak 2 25 3 35 4 45 5 55 6 65 [s] Reliable RNA fractionation with highest selectivity Total RNA was isolated from 1 7 HeLa cells using NucleoSpin mirna ( ) and two competitor kits based on phenol / chloroform lysis and extraction ( ) or phenol / chloroform extraction ( ). Equal amounts of total RNA fractions were analyzed on an Agilent Bioanalyzer (A). The NucleoSpin mirna Kit provides highest RNA yield and quality. NucleoSpin mirna allows isolation of small (B) and large (C) RNA in separate fractions in addition to the total RNA fraction (A). NucleoSpin mirna 1 / 5 / 25 74971.1 /.5 /.25 NucleoSpin mirna Plasma Mini spin kit for isolation of small RNA and DNA from plasma, serum, and exosomes n Simple and fast procedure No phenol / chloroform extraction necessary n Efficient isolation of RNA from plasma or serum samples as well as exosomes Superior recovery of small RNA > 15 nt n Optional on-column DNA digest for increased sensitivity in downstream applications Parallel DNA isolation possible if needed Reference Tan et al. Evaluation of extraction kits and RT-qPCR systems adapted to high-throughput platform for circulating mirnas Sci Rep. 215 Mar 24; 5: 943 NucleoSpin mirna Plasma 1 / 5 / 25 74981.1 /.5 /.25 6
Exosome Precipitation Solution (Serum / Plasma) (Urine) Solution for precipitation of exosomes from serum / plasma, or urine samples n Simple and fast precipitation of exosomes Avoid tedious ultracentrifugation n Flexibility in sample amount Procedure can be scaled up or down depending on sample volume n Subsequent mirna isolation possible Use NucleoSpin mirna Plasma for mirna isolation from exosomes C T (mir-16) 24 22 2 18 16 14 12 1 MN Competitor I Competitor S High mirna yields from plasma exosomes Exosomes were enriched from 25 µl plasma using Exosome Precipitation Solution from MN and competitor products. The precipitated exosomes were used for mirna isolation with NucleoSpin mirna Plasma. Quantification of mir-16 was done by quantitative realtime RT-PCR (Applied Biosystems, TaqMan MicroRNA RT Kit, hsa-mir-16 MicroRNA Assay). The results show that the use of MACHEREY-NAGEL Exosome Precipitation Solution results in highest mirna yield indicating most efficient exosome enrichment. Plasma Customer testimonial We have used the Exosome Precipitation Solution from MN to isolate exosomes from control or HCV infected sera clinical samples according to the protocol. The resulted precipitated fraction was of high yield and enriched in exosomes. The obtained fraction was pure when tested by western blotting with antibodies against HSP 9, CD 9. We are very satisfied with this product and we already recommend it to other investigators. U. Georgopoulou (PhD), Lab. Molecular Virology, Hellenic Pasteur Institute, Athens, Greece Product Pack of REF Exosome Precipitation Solution (Serum / Plasma) * 2 ml / 12 ml / 6 ml 74398.2 /.12 /.6 Exosome Precipitation Solution (Urine) * 12 ml / 2 ml / 25 ml 74399.12 /.5 /.25 *Not available in the USA NucleoSpin TriPrep Mini spin kit for parallel isolation of RNA, DNA, and proteins n Convenient one-column preparation of RNA, DNA, and proteins from unsplitted samples High quality RNA and DNA, ready to use for downstream applications n High protein yield independent of protein size, localization, modification, etc. Easy protein quantification using the MACHEREY-NAGEL Protein Quantification Assay n Complete kit with NucleoSpin Filters NucleoSpin Filters (shredders) for efficient lysis and optimized RNA recovery Reference Hansen et al. A link between inflammation and metastasis: serum amyloid A1 and A3 induce metastasis, and are targets of metastasis-inducing S1A4 Oncogene 215 Jan 22; 34 (4): 424-35 NucleoSpin TriPrep 1 / 5 / 25 74966.1 /.5 /.25 Protein Quantification Assay 5 / 25 74967.5 /.25 7
NucleoSpin RNA/Protein Mini spin kit for parallel isolation of RNA and proteins n No splitting of samples required Reliable analysis of RNA and protein levels extracted from the same sample n High RNA yield and integrity NucleoSpin Filters (shredders) and recombinant DNase for DNA-free RNA of high quality n High protein yield independent of protein size, localization, modification, etc. Easy protein quantification using the MACHEREY-NAGEL Protein Quantification Assay Reference Park et al. Targeting of mir 34a NOTCH1 Axis Reduced Breast Cancer Stemness and Chemoresistance Cancer Res., Dec 214; 74: 7573-7582 NucleoSpin RNA / Protein 1 / 5 / 25 74933.1 /.5 /.25 Protein Quantification Assay 5 / 25 74967.5 /.25 NucleoSpin RNA/DNA Buffer Set Buffer set for parallel isolation of RNA and DNA with NucleoSpin RNA kits n To be used in combination with NucleoSpin RNA kits NucleoSpin RNA, NucleoSpin RNA XS, NucleoSpin mirna, NucleoSpin RNA Blood, NucleoSpin RNA Plant, NucleoSpin RNA/Protein kits n No need to split samples (e.g., precious, indivisible samples like biopsy material) High-quality DNA and RNA from one sample, perfect for PCR, RT-PCR, real-time PCR References Eves-van den Akker et al. Identification and characterisation of a hyper-variable apoplastic effector gene family of the potato cyst nematodes PLoS Pathog. 214 Sep 25; 1 (9): E 14391 NucleoSpin RNA/DNA Buffer Set 1 74944 8
NucleoSpin RNA Blood Mini spin kit for RNA isolation from fresh and frozen whole blood n Direct total blood lysis very simple and convenient procedure (patent pending) No selective erythrocyte lysis at 4 C complete processing at room temperature n Superior RNA yield and quality from up to 4 µl whole blood Efficient on-column DNA removal for an increased sensitivity in downstream applications n Compatible with common blood collection tubes and anticoagulants E.g., EDTA, citrate, and heparin suitable for fresh and frozen whole blood samples n NucleoSpin RNA Blood Midi: For large whole blood samples Up to 26 µg high-quality RNA from 13 µl fresh or frozen whole blood samples 8 7 6 Fluorescence [RFU] 5 4 3 2 1 1 MN Blood A MN Blood B Q Blood A Q Blood B 15 2 25 3 35 Cycle number Direct lysis results in higher yields compared to selective erythrocyte lysis RNA was isolated from 4 μl blood (EDTA) from two different donors (Blood A, B) with the NucleoSpin RNA Blood kit and a kit from Competitor Q (based on selective erythrocyte lysis). For both samples, CT values are lower for NucleoSpin RNA Blood indicating a higher RNA yield. Analysis of RNA with LightCycler RT-PCR and β-actin specific primers resulted in a 73 nt amplicon. References: Guery et al. Clinical features and viral diagnosis of two cases of infection with Middle East Respiratory Syndrome coronavirus: a report of nosocomial transmission Lancet. 213 Jun 29; 381 (9885): 2265-72 NucleoSpin RNA Blood 1 / 5 742.1 /.5 NucleoSpin RNA Blood Midi 2 7421.2 NucleoSpin 8/96 RNA Blood RNA isolation from whole blood in flexible 8-well strip or proven 96-well format n Direct total blood lysis no selective erythrocyte lysis at 4 C Complete processing at RT n Compatible with common blood collection tubes and anticoagulants E.g., EDTA, citrate, and heparin suitable for fresh and frozen whole blood samples n Highly pure RNA rdnase for DNA removal included MN Wash Plate minimizes risk of cross-contamination NucleoSpin 8 RNA Blood 12 x 8 / 6 x 8 7422 /.5 NucleoSpin 96 RNA Blood 2 x 96 / 4 x 96 74225.2 /.4 9
NucleoSpin totalrna FFPE Mini spin kit for the isolation of small and large RNA from FFPE samples n Convenient paraffin removal with blue colored Paraffin Dissolver (patent pending) No xylene required n Isolate total RNA regardless of fragment size Including small RNA, such as mirna n High-quality RNA for enhanced RT-PCR performance Decrosslinking for improved RNA quality and rdnase for on-column DNA digestion 27 26 25 55 MN Excellent RT-PCR performance and most efficient gdna removal with NucleoSpin totalrna FFPE C T Values 24 23 22 21 C T Values 45 35 Q A P Large (e.g., mrna) and small (e.g., mirna) RNA was isolated from 4 x 1 μm FFPE sections of mouse brain tissue with NucleoSpin totalrna FFPE and compared to three other competitor kits (Q, A, P). (A) Quantification of mrna* and mirna** was done by qrt-pcr. Low C T values indicate high RNA yields. (B) Residual DNA was assayed by amplifying a 191 bp fragment of the mgapdh gene. The higher the C T values the lower the residual DNA. 2 *Target: 23 bp fragment of the β 2 -microglobulin gene 19 mrna mirna 25 gdna **Applied Biosystems, TaqMan MicroRNA RT Kit, hsa-mir-16 MicroRNA Assay Reference Al-Samadi et al. Increased beta 2 defensin in recurrent aphthous ulcer Oral Dis. 215 Apr; 21 (3): 292-8 NucleoSpin totalrna FFPE 1 / 5 / 25 74982.1 /.5 /.25 NucleoSpin totalrna FFPE XS Isolation of small and large RNA from very small FFPE samples n Blue colored Paraffin Dissolver (patent pending) for easy paraffin removal No xylene required n Removal of formalin crosslinking by efficient decrosslinking buffer High-quality RNA (including mirna) for improved performance in downstream applications n Special NucleoSpin XS Column for very small samples (< 5 mg tissue) Elution volume down to 5 µl for concentrated RNA NucleoSpin totalrna FFPE XS 1 / 5 / 25 74969.1 /.5 /.25 1
NucleoSpin RNA Plant Mini spin kit for the isolation of RNA from plant material n Optimized lysis procedure Two lysis buffers included choose the optimal lysis condition n Up to 7 µg ready-to-use RNA NucleoSpin Filter for efficient homogenization and reduction of viscosity n High-quality RNA rdnase for efficient DNA removal included Species Organ Yield Allium cepa (onion) Germ bud 13 μg Allium sativum (garlic) Leaf 13 μg Arabidopsis thaliana (thale cress) Leaf 15 μg Beta vulgaris (sugar beet) Leaf 17 μg Brassica napus (rapeseed) Leaf, blossom, stalk 7 9 μg Capsicum annuum (red pepper) Leaf 8 μg Cucumis melo (cucumber) Leaf 15 μg Gladiolus spec. Leaf 7 μg Hordeum vulgare (barley) Leaf 3 μg Lactuca sativa (lettuce) Leaf 4 μg Species Organ Yield Lycopersicum esculentum (tomato) Leaf 1 μg Mucor rouxii (fungus) Mycelium 6 μg Nicotiana tabacum (tobacco) Leaf, root tip, stalk, blossom 12 33 μg Secale cereale (rye) Leaf 12 μg Taraxacum officinale (dandelion) Leaf 1 μg Thymus herba-barona (thyme) Leaf 15 μg Triticum aestivum (wheat) Leaf 4 μg Viola tricolor (viola) Leaf 9 μg Zea mays (maize) Leaf 18 μg NucleoSpin RNA Plant 1 / 5 / 25 74949.1 /.5 /.25 NucleoTrap mrna Kit for the enrichment and isolation of poly(a) messenger RNA (mrna) from total RNA n High-quality mrna without DNA contamination Ready to use in all common downstream applications n Convenient and fast processing NucleoTrap Microfilters included for microcentrifugation mrna enrichment in only 3 min n Available in Mini and Midi format NucleoTrap mrna Mini: for up to 25 μg total RNA as starting material NucleoTrap mrna Midi: for up to 1, μg total RNA as starting material NucleoTrap mrna Mini 12 74655 NucleoTrap mrna Midi 12 74656 11
RNA from cells and tissue Preps / Pack of REF NucleoSpin RNA 1 / 5 / 25 74955.1 /.5 /.25 NucleoSpin RNA Plus 1 / 5 / 25 74984.1 /.5 /.25 NucleoZOL 2 ml 7444.2 NucleoSpin RNA Set for NucleoZOL 1 / 5 7446.1 /.5 NucleoSpin RNA XS 1 / 5 / 25 7492.1 /.5 /.25 NucleoSpin RNA Midi 2 74962.2 NucleoSpin 8 RNA 12 x 8 / 6 x 8 74698 /.5 NucleoSpin 8 RNA Core Kit 48 x 8 74465.4 NucleoSpin 96 RNA 2 x 96 / 4 x 96 / 24 x 96 7479.2 /.4 /.24 NucleoSpin 96 RNA Core Kit 4 x 96 74466.4 NucleoMag RNA 1 x 96 / 4 x 96 74435.1 /.4 MicroRNA Preps / Pack of REF NucleoSpin mirna 1 / 5 / 25 74971.1 /.5 /.25 NucleoSpin mirna Plasma 1 / 5 / 25 74981.1 /.5 /.25 Exosome Precipitation Solution (Serum / Plasma)* 2 ml / 12 ml / 6 ml 74398.2 /.12 /.6 Exosome Precipitation Solution (Urine)* 12 ml / 2 ml / 25 ml 74399.12 /.5 /.25 RNA, DNA, and protein Preps REF NucleoSpin TriPrep 1 / 5 / 25 74966.1 /.5 /.25 NucleoSpin RNA/Protein 1 / 5 / 25 74933.1 /.5 /.25 NucleoSpin RNA/DNA Buffer Set 1 74944 RNA from blood Preps REF NucleoSpin RNA Blood 1 / 5 742.1 /.5 NucleoSpin RNA Blood Midi 2 7421.2 NucleoSpin 8 RNA Blood 12 x 8 / 6 x 8 7422 /.5 NucleoSpin 96 RNA Blood 2 x 96 / 4 x 96 74225.2 /.4 Small and large RNA from FFPE samples Preps REF NucleoSpin totalrna FFPE 1 / 5 / 25 74982.1 /.5 /.25 NucleoSpin totalrna FFPE XS 1 / 5 / 25 74969.1 /.5 /.25 RNA from plant Preps REF NucleoSpin RNA Plant 1 / 5 / 25 74949.1/.5/.25 Poly(A) mrna from total RNA Preps REF NucleoTrap mrna Mini 12 74655 NucleoTrap mrna Mini 12 74656 RNA clean-up Preps REF NucleoSpin RNA Clean-up 1 / 5 / 25 74948.1 /.5 /.25 NucleoSpin RNA Clean-up XS 1 / 5 / 25 7493.1 /.5 /.25 Trademarks: PAXgene registered trademark by PreAnalytiX GmbH, Agilent Bioanalyzer registered trademark by Agilent Technologies, LightCycler registered trademark by Roche Group, TaqMan registered trademark by Life Technologies Corporation *Not available in the USA Fly_dragonfly, HYUNGKEUN, Paul Binet fotolia.com KATEN391 RNA Guide en4 / 2 / / 3.217 DD Printed in Germany MACHEREY-NAGEL GmbH & Co. KG Neumann-Neander-Str. 6 8 52355 Düren Germany DE / International: CH: FR: Tel.: +49 24 21 969- Fax: +49 24 21 969-199 E-mail: info@mn-net.com Tel.: +41 62 388 55 Fax: +41 62 388 55 5 E-mail: sales-ch@mn-net.com Tel.: +33 388 68 22 68 Fax: +33 388 51 76 88 E-mail: sales-fr@mn-net.com US: Tel.: +1 484 821 984 Fax: +1 484 821 1272 E-mail: sales-us@mn-net.com